Dear all,
I am running some gel filtration FPLC analysis in an AKTA Pure system.
There's an issue, the pressure in the system with no column attached is already ~0.3 MPa, which is very high considering the column limit of 0.5 MPa. (The system I work with measures the pre-column pressure (preC) in R9 at the pump system outlet).
"what should be the pressure in the system with no column attached?"
(I already did some mantainance and checked/replaced the tubing with no results.
However, before going on with the cleaning I'd like to know what pressure I should expect without column)
Thanks in advance,
Andrea
The system pressure w/o a column inline will depend on the mobile phase solution viscosity, method used, specific plumbing on YOUR instrument and the flow rate. No one can provide an exact value as each system will be different. You must determine what yours is, using your application method and instrument.
If the pressure is abnormally high, please troubleshoot the flow path step-by-step, checking all connections and fittings. Have someone local who is experienced in using your AKTA system assist you in this process as the web is not very useful for these types of issues (onsite assistance is required).
The system pressure limit is much higher than the column pressure limit. According to Cytiva, the system pressure limit for an AKTA Pure is safe up to 20 MPa. As William has already explained, pressure is dependent on many factors such as viscosity, so 0.3 MPa seems to be the running pressure of your current buffer/set up. When you run a method, as long as the pressure limit is set to 0.5 MPa for the column, it won't exceed this. During your method, pressure is likely to remain at 0.3 MPa unless a blockage occurs or you set the flow rate too high.
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